Figure 7.

H₂O₂ exposure induced ferroptosis in L02 cells. (a) Relative cell viability was confirmed by the CCK-8 assay (n = 6). (b) L02 cells underwent ferroptosis-specific morphologic phenotype (cells shrunk at first and eventually exhibiting a balloon-type) after being treated with H₂O₂ or Erastin. The blue and red arrows indicate shrunken and balloon phenotypes, respectively ((a) normal L02 cells. (b) L02 cells treated with H₂O₂. (c) L02 cells co-treated with Fer-1 and H₂O₂. (d) L02 cells exposed to Erastin). Original magnification ×200. (c) Cells were stained with 5 μM BODIPY® 581/591 C11 to detect the content of cellular lipid peroxide through flow cytometry (n = 3). (d and e) Cellular content MDA and total GSH were measured, respectively (n = 3). (f) Protein expression of Nrf2 was elevated after cells were treated with H₂O₂ or Erastin compared with normal L02 cells (n = 3). (g) The mRNA expression of HO-1 and NQO1 was elevated in response to H₂O₂ and Erastin treatment (n = 3). ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001. CCK-8 = Cell Counting Kit-8; Fer-1 = ferrostatin-1; GSH = glutathione; HO-1 = heme oxygenase-1; MDA = malondialdehyde; NQO1 = NAD(P) H quinone dehydrogenase, quinone 1.